Effect of CCK pretreatment on the CCK sensitivity of rat polymodal gastric vagal afferent in vitro

To prevent the blood-borne interference and reflex actions via neighboring organs and the central nervous system, the study was conducted in an in vitro isolated stomach-gastric vagus nerve preparation obtained from overnight-fasted, urethan-anesthetized rats. Afferent unit action potentials were recorded from the gastric branch of the vagus nerve. The left gastric artery was catheterized for intra-arterial injection. In vitro we found that 1) 55/70 gastric vagal afferents (GVAs) were polymodal, responding to CCK-8 and mechanical stimuli, 13 were mechanoreceptive, and 2 were CCK-responsive; 2) sequential or randomized intra-arterial injections of CCK-8 (0.1–200 pmol) dose-dependently increased firing rate and reached the peak rate at 100 pmol; 3) the action was suppressed by CCK-A (Devazepide) but not by CCK-B (L-365,260) receptor antagonist; 4) neither antagonist blocked the mechanosensitivity of GVA fibers. These results are consistent with corresponding in vivo well-documented findings. Histological data indicate that the layered structure of the stomach wall was preserved in vitro for 6–8 h. Based on these results, it seems reasonable to use the in vitro preparation for conducting a study that is usually difficult to be performed in vivo. For instance, because there was no blood supply in vitro, the composition of the interstitial fluid, i.e., the ambient nerve terminals, can be better controlled and influenced by intra-arterial injection of a defined solution. Here we report that acutely changing the ambient CCK level by a conditioning stimulus (a preceding intra-arterial injection of increasing doses of CCK-8) reduced the CCK sensitivity of GVA terminals to a subsequent test stimulus (a constant dose of CCK-8 intra-arterial injection).

Identification of muscle afferents subserving sensation of deep pain in humans

1. Intraneural microstimulation (INMS) and microneurography were used in combination to stimulate and record from muscle nociceptor primary afferent fibers of the common peroneal nerve of healthy volunteers. When pain evoked by INMS was projected to muscle, afferent activity could be evoked by innocuous and noxious pressure applied within the projected painful area. Conduction velocity of single fibers was determined by stimulating the receptive fields (RFs) electrically via needle electrodes inserted into the RF and measuring conduction latency and distance between the RF and recording electrode. 2. Pain projected to muscle during INMS trains 5–10 s in duration at threshold intensity for pain sensation was typically described as cramping and was well localized. Subjects mapped the area of the painful projected field (PF) over the skin using a pointer. 3. Fourteen slowly adaping mechanoreceptors with RF in muscle and with moderate to high receptor threshold were identified within or near the painful PF. Conduction velocities were in the range of Group III (n = 8) and Group IV (n = 6) fibers. Mean RF areas of Group III and Group IV afferents, determined by applying pressure percutaneously, were 2.71 +/- 1.14 (SE) cm2 and 3.40 +/- 1.08 (SE) cm2, respectively. Only one Group III afferent unit exhibited spontaneous activity (< 1 Hz). 4. One additional high-threshold mechanoreceptor was identified, with its RF located in the extensor tendon at the base of the big toe. This fiber had a conduction velocity of 32 m/s. During INMS, a well-localized sharp pain was projected to the tendon.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential activation and classification of cutaneous afferents in the rat

1. A total of 312 cutaneous afferent units identified in the rat foot as belonging to one of nine major types of sensory receptors were included in the present study. A natural stimulus set was defined to differentiate optimally among those receptor types according to the distinguishing response patterns that it produced. It included air puffs, 30- and 300-Hz sinusoids, 200-mN force indentation of the skin, 1.2- and 6-N compressions of a skin fold, cooling the skin by 5 and 20 degrees C, warming by 5 degrees C, and heating by 15 degrees C. 2. The responses to predefined stimuli of 188 units were subjected to multivariate statistical analyses. The responses of an individual unit were measured as the number of impulses evoked by 10 stimuli, each lasting 10 s. Additionally, the number of impulses occurring for 5 s after withdrawal of a 200-mN indentation (1 of the 10 stimuli) was counted. 3. In discriminant analysis, the 11 stimulus variables predicted fairly correctly the grouping of afferent units into nine predetermined receptor categories (175 of 188, 93.1%), indicating a powerful ability to discriminate among different receptor types. Using hierarchical cluster analysis, afferent unit data described by 11 variables were divided into clusters that well represented prior receptor categories (170 of 188, 90.4%), suggesting the reliable application of this procedure to the classification of newly recorded cutaneous sensory receptors. 4. Eleven variables were then reduced to 7 on the basis of the results of factor analysis (95% of variance accounted for). The seven variables corresponded to 1.2-N compression, heating the skin by 15 degrees C, cooling the skin by 20 degrees C, 30- and 300-Hz sinusoids, withdrawal of a 200-mN indentation, and air puffs. 5. The seven selected variables correctly assigned afferent units into five modality-based categories in the discriminant solution (177 of 188, 94.1%). In the cluster solution, afferent units described by the seven selected variables were divided into clusters, most of whose members were modality specific (176 of 188, 93.6%). 6. The results indicate that cutaneous receptors can be divided into modality-specific groups according to similarities in their responses to seven stimulus variables. It is proposed that the stimulus set developed here and multivariate statistical methods can be used as powerful tools for the functional classification of central somatosensory neurons.

Hypothalamic modulation of baroreceptor afferent unit activity

Unit responses to sinus nerve stimulation were recorded in the medulla. A conditioning stimulus to the posterior hypothalamus produced inhibition of 65% of unit responses to sinus nerve stimulation as early as 7 ms and extending as long as 790 ms after conditioning; 50% recovered after 300 ms. Unit responses to hypothalamic stimulation alone were also recorded in the medulla, some in the same loci as other unit responses to sinus nerve stimulation. They could be activated by contralateral as well as ipsilateral hypothalmic stimulation and showed recurrent bursts of firing over a 1,000-ms poststimulus interval. Evoked potentials and unit responses were recorded in the posterior hypothalamus, some occurring within 10--20 ms poststimulation of the sinus nerve, indicating that baroreceptor information is ascending in a time sufficiently short to involve the hypothalamus in reflex regulation of blood pressure as well as more generalized homeostatic responses which include the cardiovascular system.